Structural studies of the endoglycosidase H-resistant oligosaccharides present on human beta-glucuronidase.
Human beta-glucuronidase bears 3-4 oligosaccharide moieties/subunit of Mr = 75,000. We have previously characterized the endoglycosidase H-releasable oligosaccharides of this enzyme including those which are phosphorylated and involved in targeting to lysosomes. In this study, we report the characterization of the endoglycosidase H-resistant oligosaccharides which were released from beta-glucuronidase with anhydrous hydrazine. Approximately 65% of the hydrazine-released oligosaccharides are of the high mannose type, with the predominant species containing 9 mannose residues. The remaining oligosaccharides appear to originate from incomplete complex oligosaccharides. Their basic structures are Man alpha 1,6Man beta 1,4Glc-NAc beta 1,4GlcNAcol, and Man alpha 1,3[Man alpha 1,6]Man beta 1,4Glc-NAc beta 1,4GlcNAcol with roughly half of each species containing an additional fucose linked alpha 1,6 to the N-acetylglucosaminitol (GlcNAcol) residue. The small amount of complex oligosaccharide present bearing 1 sialic acid was heterogeneous in nature with incompletion of the nonsialylated branch. In addition, there was a minor specie of high mannose-type oligosaccharide bearing 5 mannose residues with an alpha 1,6-linked fucose on the GlcNAcol. This structure was not expected since high mannose-type oligosaccharides have been reported to not be substrates for the alpha 1,6-fucosyl transferase.
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D005966:Glucuronidase; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D009844:Oligosaccharides / Q000032:analysis*
D046911:Macromolecular Substances; D009844:Oligosaccharides; D006026:Glycoside Hydrolases; D005966:Glucuronidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase